INFORMATION
| Full Name | Luteinizing Hormone Releasing Hormone |
|---|---|
| Synonym | LHRH/GnRH |
| Assay Type | Competitive |
| Reactivity | Porcine |
| Range | 15.63-1000pg/mL |
| Sensitivity | 9.38pg/mL |
| Sample Type | Serum, plasma and other biological fluids |
| Sample Volume | 50μL |
| Detection Wavelength | OD450 |
| Reaction Duration | 1.5h |
| Transportation Temperature | 2-8°C |
| Specificity | Specifically binds with LHRH/GnRH, no obvious cross reaction with other analogues. |
| Microplate | 96-wells plate breakable into 12 x 8 wells strip |
ELISA KIT COMPONENTS
Upon receipt, unpack promptly and store as recommended in the instructions.
| Components | Specifications | Storage and Notes |
|---|---|---|
| Micro Plate | 96T: 8 wells×12 strips 48T: 8 wells×6 strips | Unopened: -20°C, 12 months Unused: Put it back in the aluminum foil bag and seal it, store it at -20°C. |
| Reference Standard | 96T: 2 vials 48T: 1 vial | Unopened: -20°C, 12 months Please use freshly dissolved standards for each experiment. Discard any unused standards after dissolution. |
| Biotinylated Detection Ab Concentrate (100×) | 96T: 120μL×1 vial 48T: 60μL×1 vial | Unopened: -20°C, 12 months Unused: Please seal the concentrate and store it at -20°C, and discard the working solution. |
| HRP Conjugate Concentrate (100×) | 96T: 120μL×1 vial 48T: 60μL×1 vial | Unopened: -20°C(Protect from light), 12 months Unused: Please seal the concentrate and store it at -20°C, and discard the working solution. |
| Biotinylated Detection Ab Diluent | 14mL×1 | 2-8℃, 12 months |
| HRP Conjugate Diluent | 14mL×1 | 2-8℃, 12 months |
| Reference Standard & Sample Diluent | 20mL×1 | 2-8℃, 12 months |
| Washing Buffer Concentrate (25×) | 30mL×1 | 2-8℃, 12 months |
| Substrate Reagent(TMB) | 10mL×1 | 2-8°C(Protect from light),12 months |
| Stop Solution | 7mL×1 | 2-8°C/Room temperature |
REQUIRED INSTRUMENTS AND REAGENTS
- Microplate reader (450nm wavelength filter).
- Incubator capable of maintaining 37℃.
- 1.5mL EP tube, absorbent paper.
- Pipette and disposable tip: 0.5-10μL, 2-20μL, 20-200μL, 200-1000μL.
- Double distilled water or deionized water.
ASSAY PROCEDURE SUMMARY
STANDARD CURVE
| STD.(pg/mL) | OD-1 | OD-2 | Average |
|---|---|---|---|
| 0 | 2.252 | 2.27 | 2.261 |
| 15.63 | 1.841 | 1.851 | 1.846 |
| 31.25 | 1.542 | 1.54 | 1.541 |
| 62.5 | 1.163 | 1.163 | 1.163 |
| 125 | 0.796 | 0.816 | 0.806 |
| 250 | 0.547 | 0.543 | 0.545 |
| 500 | 0.368 | 0.408 | 0.388 |
| 1000 | 0.288 | 0.32 | 0.304 |
RECOVERY
The recovery of Porcine LHRH/GnRH spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.
LINEARITY
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Porcine LHRH/GnRH in various matrices were diluted with the Reference Standard & Sample Diluent to produce samples with values within the dynamic range of the assay.
PRECISION(%)
Intra-Assay Precision (Precision within an assay): Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays): Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
| Item | Intra-assay Precision | Inter-assay Precision | ||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 49.06 | 86.54 | 424.34 | 50.48 | 82.47 | 450.11 |
| Standard deviation | 3.12 | 3.67 | 12.98 | 2.6 | 3.99 | 22.37 |
| CV(%) | 6.36 | 4.24 | 3.06 | 5.16 | 4.84 | 4.97 |
STABILITY
Perform stability testing of the kit at 37°C and 2-8°C and obtain relevant data.
| ELISA kit(n=5) | 37°C for 1 month | 2-8°C for 6 months |
|---|---|---|
| Average(%) | 80 | 95-100 |
